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30

Bases on chemical composition of cellular components.

Decolorizing agent

31

Contrasting color to that of the primary stain

Counterstain

32

Differential staining procedure developed by Hans Christian Gram in 1884

Gram staining

33

Primary stain crystal violet (basic purple dye) applied to heat stain

Gram staining step 1

34

After washing primary stain iodine (mordant) is applied that will make crystal violent complex
Bacteria at this point appear dark violet or purple

Gram staining step 2

35

Alcohol, alcohol acetone solution applied as decoloring agent and removes purple color from Gram negative bacteria
** Gram Positive will retain purple color

Gram Staining step 3

36

Alcohol rinsed and the slide is then stained with Counterstain Safranin(basic red dye)

Gram Staining step 4

37

Have thicker layer if Peptidoglycan in cell wall. CV-1 complex is not washed by alcohol wash. They retain the CV-1 complex and remain purple

Gram positive bacteria

38

Have thinner layer of Peptidoglycan and also have layer of lipopolysachharide as part of their cell wall.
Alcohol disrupts layer and CV-1 complex is washed through
***will remain colorless unless counter stain Safranin applied, become pink

Gram negative

39

Penicillins
Cephalosporins

*cannot penetrate lipopolysachharide layer of gram negative**

Beta Lactams

40

Uses to distinguish Myobacterium species and some species of Nocardia
Bind strongly to waxy material of the cell wall

Acid fast staining

41

Carbol fushion(red dye) applied to fixed smear








Acid fast staining step 1

42

After cooling & washing smear washed with alcohol decolorizer
*Non acid fast lose stain
**Acid fast retain dye, more soluble in lipid cell wall than in acid alcohol

Acid fast staining step 2

43

Counter stain methylene blue is used to stain non acid fast bacteria blue

Acid fast staining step 3

44

Schaeffer-Fulton stain
1. Heat fixed smear
2. Primary stain Malachite green applied, heated (stain wall)
3. Washed to remove excess
4. Counter stain Safranin applied to stain other parts pink

Endospore special stains

45

Determines microbes virulence
Gelatinous covering that is soluble in water.
India Ink or Nigrosin stains background dark
Safranin used to stain cells
Capsules look like Halos around stained cells

Capsule staining

46

Mordant used to build diameter
When stained with Carbolfuchsin becomes visible under light microscope.
Number and location used in diagnosis

Flagella Staining

47

4x

Scanning

48

10x

Low power

49

40-45x

High power

50

90-100x

Oil immersion

51

Ocular lens

10x

52

Ocular lens x objective lens

Total magnification

53

Any kind of microscope that uses visible light to observe specimen

Are compound microscope

Light microscope

54

Microscope remains in focus even objectives are changed

Parfocal

55

Acid fast color

Red

56

Non acid fast color

Blue